gateway bp clonase ii enzyme mix  (Thermo Fisher)


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    Structured Review

    Thermo Fisher gateway bp clonase ii enzyme mix
    Gateway Bp Clonase Ii Enzyme Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 155 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gateway bp clonase ii enzyme mix/product/Thermo Fisher
    Average 99 stars, based on 155 article reviews
    Price from $9.99 to $1999.99
    gateway bp clonase ii enzyme mix - by Bioz Stars, 2020-02
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Phytoplasma SAP11 effector destabilization of TCP transcription factors differentially impact development and defence of Arabidopsis versus maize
    Article Snippet: .. The GFP-sequence was amplified from pUBN-GFP-DEST with the gene-specific primers STOP-GFP forward and reverse , cloned into pDONR207 with the Gateway BP Clonase II Enzyme Mix (Invitrogen) and transferred to pUBN-GFP-DEST using the Gateway LR Clonase II Enzyme Mix (Invitrogen). .. Isolation and transformation of Arabidopsis and maize protoplasts were performed as described by [ ].

    Article Title: Host‐induced gene silencing compromises Verticillium wilt in tomato and Arabidopsis
    Article Snippet: .. The DNA fragments were cloned into pDONR207 using the Gateway® BP Clonase® II Enzyme Mix (Invitrogen, Carlsbad, CA, USA) to generate entry vectors, and all the entry vectors were verified by DNA sequencing (Eurofins Genomics, Ebersberg, Germany). .. Subsequently, the entry vector pDONR207 carrying the Ave1 fragment was transferred to TRV2 and pFAST R03 to generate constructs TRV2::Ave1 and pFAST R03_Ave1 (Fig. ), respectively, and pDONR207 entry vectors carrying the NLP1 or Sge1 fragment were recombined into pTRV2 and pHellsgate 12 to generate constructs TRV2::NLP1 , TRV2::Sge1 , pHellsgate 12_NLP1 and pHellsgate 12_Sge1 (Fig. ) using Gateway® LR Clonase® II Enzyme Mix (Invitrogen).

    Article Title: Cytokine exocytosis and JAK/STAT activation in the Drosophila ovary requires the vesicle trafficking regulator α-Snap
    Article Snippet: The coding sequence – excluding the stop codon – to allow expression of the HA tag was amplified using primers that contained attB sites following the GateWay cloning protocol by Invitrogen/ThermoFisher. .. The BP reaction was carried out using a pDNOR221 vector (Invitrogen, 12536-017) and Gateway BP Clonase II enzyme mix (Invitrogen, 11789020) followed by heat shock-induced transformation into One Shot OmniMax 2T1 competent E. coli cells (Invitrogen, C854003).

    Article Title: IRAK3 modulates downstream innate immune signalling through its guanylate cyclase activity
    Article Snippet: .. Generation of IRAK3 and IRAK4 constructs The IRAK3 gene in pBluescriptR (MHS1010-9204142, clone ID 30335802) was cloned into pDONR207 using Gateway® BP Clonase™ II Enzyme Mix (Invitrogen, Thermo Scientific USA). .. Five primers were designed to make four constructs; two forward primers, each containing the Kozak sequence and three reverse primers to either incorporate a Myc tag or a STOP codon (Supplementary Table ) and these were generated by PCR.

    Article Title: Differential Roles of the Thylakoid Lumenal Deg Protease Homologs in Chloroplast Proteostasis
    Article Snippet: .. The PCR products were recombined with the pDONR221 vector using Gateway BP-Clonase II Enzyme Mix (Invitrogen), yielding entry clones. .. Plasmids expressing the Deg genomic DNA with C-terminal His tags were constructed by performing LR-trans reactions between the pDONR221-Deg entry vectors and the pGWB407/pGWB607 destination vectors using the Gateway LR-Clonase II Enzyme Mix (Invitrogen).

    Article Title: A dispensable paralog of succinate dehydrogenase subunit C mediates standing resistance towards a subclass of SDHI fungicides in Zymoseptoria tritici
    Article Snippet: .. To generate the SDHC1 and SDHC3 KO mutants, 5’ upstream regions of 1000bp and 2074 bp and 3’ downstream regions of 914bp and 1313bp for ZtSDHC1 and ZtSDHC3 respectively were PCR-amplified from genomic DNA of IPO323 or 06STD024 strains and the fragments cloned by BP cloning using Gateway BP Clonase II Enzyme Mix (Invitrogen) into pDONR-P4-P1R (upstream regions) or pDONR-P2R-P3 (downstream regions) ( for oligos). .. These 5’ and 3’ gene locus-paired entry plasmids were then combined with the pENTR221-TrpChyg described previously [ ] and pNOV2114_gateway for multisite gateway LR cloning using Gateway LR Clonase II Plus enzyme Mix (Invitrogen) following provider’s instructions.

    Article Title: Phytoplasma SAP11 effector destabilization of TCP transcription factors differentially impact development and defence of Arabidopsis versus maize
    Article Snippet: .. The fragments were further amplified with attB1 and attB2 adapter primers and cloned into pDONR207 with Gateway BP Clonase II Enzyme Mix (Invitrogen, Carlsbad, USA). ..

    Article Title: Haustoria – arsenals during the interaction between wheat and Puccinia striiformis f. sp. tritici
    Article Snippet: .. For pEDV6, the coding sequence (without a signal peptide) was cloned and recombined in pDONR221 following the instructions of the Gateway BP clonase II enzyme mix and Gateway LR clonase enzyme mix II (Invitrogen, Carlsbad, CA, USA). ..

    Amplification:

    Article Title: Phytoplasma SAP11 effector destabilization of TCP transcription factors differentially impact development and defence of Arabidopsis versus maize
    Article Snippet: .. The GFP-sequence was amplified from pUBN-GFP-DEST with the gene-specific primers STOP-GFP forward and reverse , cloned into pDONR207 with the Gateway BP Clonase II Enzyme Mix (Invitrogen) and transferred to pUBN-GFP-DEST using the Gateway LR Clonase II Enzyme Mix (Invitrogen). .. Isolation and transformation of Arabidopsis and maize protoplasts were performed as described by [ ].

    Article Title: Host‐induced gene silencing compromises Verticillium wilt in tomato and Arabidopsis
    Article Snippet: Selected DNA fragments were amplified by PCR from the corresponding plasmids using the gene‐specific primers listed in Table S1 (see Supporting Information). .. The DNA fragments were cloned into pDONR207 using the Gateway® BP Clonase® II Enzyme Mix (Invitrogen, Carlsbad, CA, USA) to generate entry vectors, and all the entry vectors were verified by DNA sequencing (Eurofins Genomics, Ebersberg, Germany).

    Article Title: Cytokine exocytosis and JAK/STAT activation in the Drosophila ovary requires the vesicle trafficking regulator α-Snap
    Article Snippet: The coding sequence – excluding the stop codon – to allow expression of the HA tag was amplified using primers that contained attB sites following the GateWay cloning protocol by Invitrogen/ThermoFisher. .. The BP reaction was carried out using a pDNOR221 vector (Invitrogen, 12536-017) and Gateway BP Clonase II enzyme mix (Invitrogen, 11789020) followed by heat shock-induced transformation into One Shot OmniMax 2T1 competent E. coli cells (Invitrogen, C854003).

    Article Title: Chemical Screening Pipeline for Identification of Specific Plant Autophagy Modulators 1
    Article Snippet: Fluc and Rluc coding sequences were amplified using the primer pairs PB31/PB32 and PB33/PB34 and vectors pRD400 35S::Fluc and pRD400 35S::Rluc , respectively. .. The synthetic sequence of attB-flanked SV40 NLS (Eurofins Scientific) was recombined with the Gateway pDONRTM/Zeo vector (ThermoFisher) using Gateway BP Clonase II enzyme mix (ThermoFisher), producing entry clone CC3.

    Article Title: Differential Roles of the Thylakoid Lumenal Deg Protease Homologs in Chloroplast Proteostasis
    Article Snippet: The Deg1 , Deg5 , and Deg8 promoter and coding sequences were amplified from wild-type genomic DNA with attB . .. The PCR products were recombined with the pDONR221 vector using Gateway BP-Clonase II Enzyme Mix (Invitrogen), yielding entry clones.

    Article Title: Chemical Screening Pipeline for Identification of Specific Plant Autophagy Modulators 1
    Article Snippet: .. The obtained attB-flanked amplicon was then recombined with the Gateway pDONR/Zeo vector (ThermoFisher) using Gateway BP Clonase II enzyme mix (ThermoFisher). .. Fluc and Rluc coding sequences were amplified using the primer pairs PB31/PB32 and PB33/PB34 and vectors pRD400 35S::Fluc and pRD400 35S::Rluc , respectively.

    Article Title: Phytoplasma SAP11 effector destabilization of TCP transcription factors differentially impact development and defence of Arabidopsis versus maize
    Article Snippet: .. The fragments were further amplified with attB1 and attB2 adapter primers and cloned into pDONR207 with Gateway BP Clonase II Enzyme Mix (Invitrogen, Carlsbad, USA). ..

    Article Title: Complex Iron Uptake by the Putrebactin-Mediated and Feo Systems in Shewanella oneidensis
    Article Snippet: In brief, two fragments flanking the genes of interest were amplified by PCR with outside primers (O; ) containing attB and gene-specific sequences and with inside primers (I; ) containing complementary sequences and gene-specific sequences and were then linked by a second round of PCR with outside primers. .. The fused fragments were introduced into plasmid pHGM01 using Gateway BP clonase II enzyme mix (Invitrogen) according to the manufacturer's instructions.

    Article Title: Haustoria – arsenals during the interaction between wheat and Puccinia striiformis f. sp. tritici
    Article Snippet: For pGR106, we amplified HASP candidate genes from the cDNA of Suwon 11 infected with CYR31, digested them with enzymes Cla I, Sal I or Smal I, and cloned into pGR106. .. For pEDV6, the coding sequence (without a signal peptide) was cloned and recombined in pDONR221 following the instructions of the Gateway BP clonase II enzyme mix and Gateway LR clonase enzyme mix II (Invitrogen, Carlsbad, CA, USA).

    Electroporation:

    Article Title: Host‐induced gene silencing compromises Verticillium wilt in tomato and Arabidopsis
    Article Snippet: The DNA fragments were cloned into pDONR207 using the Gateway® BP Clonase® II Enzyme Mix (Invitrogen, Carlsbad, CA, USA) to generate entry vectors, and all the entry vectors were verified by DNA sequencing (Eurofins Genomics, Ebersberg, Germany). .. All constructs were transformed to Agrobacterium tumefaciens strain GV3101 (pMP90) by electroporation.

    Mutagenesis:

    Article Title: Differential Roles of the Thylakoid Lumenal Deg Protease Homologs in Chloroplast Proteostasis
    Article Snippet: Paragraph title: Generation of Mutant and Transgenic Lines, Genotyping, and Expression Analysis ... The PCR products were recombined with the pDONR221 vector using Gateway BP-Clonase II Enzyme Mix (Invitrogen), yielding entry clones.

    Article Title: Complex Iron Uptake by the Putrebactin-Mediated and Feo Systems in Shewanella oneidensis
    Article Snippet: Paragraph title: In-frame mutant construction and complementation. ... The fused fragments were introduced into plasmid pHGM01 using Gateway BP clonase II enzyme mix (Invitrogen) according to the manufacturer's instructions.

    Isolation:

    Article Title: Phytoplasma SAP11 effector destabilization of TCP transcription factors differentially impact development and defence of Arabidopsis versus maize
    Article Snippet: The GFP-sequence was amplified from pUBN-GFP-DEST with the gene-specific primers STOP-GFP forward and reverse , cloned into pDONR207 with the Gateway BP Clonase II Enzyme Mix (Invitrogen) and transferred to pUBN-GFP-DEST using the Gateway LR Clonase II Enzyme Mix (Invitrogen). .. Isolation and transformation of Arabidopsis and maize protoplasts were performed as described by [ ].

    Article Title: Differential Roles of the Thylakoid Lumenal Deg Protease Homologs in Chloroplast Proteostasis
    Article Snippet: Single mutants were backcrossed to the wild-type plants three times to remove additional mutations, and deg1 homozygous plants for the T-DNA insertions (identified by PCR analyses) were isolated. .. The PCR products were recombined with the pDONR221 vector using Gateway BP-Clonase II Enzyme Mix (Invitrogen), yielding entry clones.

    Synthesized:

    Article Title: Phytoplasma SAP11 effector destabilization of TCP transcription factors differentially impact development and defence of Arabidopsis versus maize
    Article Snippet: The fragments were further amplified with attB1 and attB2 adapter primers and cloned into pDONR207 with Gateway BP Clonase II Enzyme Mix (Invitrogen, Carlsbad, USA). .. A codon-optimized version of SAP11MBSP without the sequence corresponding to the signal peptide and DNA sequences corresponding to the TCP domains of ZmTCP9 , AtTCP12 , AtTCP18 and the AtTCP2 and SAP11 chimeras were gene synthesized by Genscript (New Jersey, USA) with Gateway compatible attL1 and attL2 attachment sites ( and Tables) and provided in pMS (Genscript).

    Construct:

    Article Title: Host‐induced gene silencing compromises Verticillium wilt in tomato and Arabidopsis
    Article Snippet: Paragraph title: Generation of the constructs ... The DNA fragments were cloned into pDONR207 using the Gateway® BP Clonase® II Enzyme Mix (Invitrogen, Carlsbad, CA, USA) to generate entry vectors, and all the entry vectors were verified by DNA sequencing (Eurofins Genomics, Ebersberg, Germany).

    Article Title: IRAK3 modulates downstream innate immune signalling through its guanylate cyclase activity
    Article Snippet: .. Generation of IRAK3 and IRAK4 constructs The IRAK3 gene in pBluescriptR (MHS1010-9204142, clone ID 30335802) was cloned into pDONR207 using Gateway® BP Clonase™ II Enzyme Mix (Invitrogen, Thermo Scientific USA). .. Five primers were designed to make four constructs; two forward primers, each containing the Kozak sequence and three reverse primers to either incorporate a Myc tag or a STOP codon (Supplementary Table ) and these were generated by PCR.

    Article Title: Chemical Screening Pipeline for Identification of Specific Plant Autophagy Modulators 1
    Article Snippet: Paragraph title: Genetic Constructs ... The synthetic sequence of attB-flanked SV40 NLS (Eurofins Scientific) was recombined with the Gateway pDONRTM/Zeo vector (ThermoFisher) using Gateway BP Clonase II enzyme mix (ThermoFisher), producing entry clone CC3.

    Article Title: Differential Roles of the Thylakoid Lumenal Deg Protease Homologs in Chloroplast Proteostasis
    Article Snippet: The PCR products were recombined with the pDONR221 vector using Gateway BP-Clonase II Enzyme Mix (Invitrogen), yielding entry clones. .. Plasmids expressing the Deg genomic DNA with C-terminal His tags were constructed by performing LR-trans reactions between the pDONR221-Deg entry vectors and the pGWB407/pGWB607 destination vectors using the Gateway LR-Clonase II Enzyme Mix (Invitrogen).

    Article Title: A dispensable paralog of succinate dehydrogenase subunit C mediates standing resistance towards a subclass of SDHI fungicides in Zymoseptoria tritici
    Article Snippet: Production of Z . tritici transformants The multisite binary pNOV2114_gateway and pNOV2114 Hyg _gateway (3-way) vectors were used to generate the different transformation constructs [ ]. .. To generate the SDHC1 and SDHC3 KO mutants, 5’ upstream regions of 1000bp and 2074 bp and 3’ downstream regions of 914bp and 1313bp for ZtSDHC1 and ZtSDHC3 respectively were PCR-amplified from genomic DNA of IPO323 or 06STD024 strains and the fragments cloned by BP cloning using Gateway BP Clonase II Enzyme Mix (Invitrogen) into pDONR-P4-P1R (upstream regions) or pDONR-P2R-P3 (downstream regions) ( for oligos).

    Article Title: Phytoplasma SAP11 effector destabilization of TCP transcription factors differentially impact development and defence of Arabidopsis versus maize
    Article Snippet: Generation of Gateway compatible entry clones We generated Gateway compatible entry clones for all experiments, except for the constructs to transform maize. .. The fragments were further amplified with attB1 and attB2 adapter primers and cloned into pDONR207 with Gateway BP Clonase II Enzyme Mix (Invitrogen, Carlsbad, USA).

    Article Title: Complex Iron Uptake by the Putrebactin-Mediated and Feo Systems in Shewanella oneidensis
    Article Snippet: In-frame deletion strains were constructed using the att -based fusion PCR method as described previously ( ). .. The fused fragments were introduced into plasmid pHGM01 using Gateway BP clonase II enzyme mix (Invitrogen) according to the manufacturer's instructions.

    Purification:

    Article Title: IRAK3 modulates downstream innate immune signalling through its guanylate cyclase activity
    Article Snippet: Generation of IRAK3 and IRAK4 constructs The IRAK3 gene in pBluescriptR (MHS1010-9204142, clone ID 30335802) was cloned into pDONR207 using Gateway® BP Clonase™ II Enzyme Mix (Invitrogen, Thermo Scientific USA). .. Plasmids were purified using PureLink® Quick Plasmid Miniprep Kit (Thermo Fisher Scientific, USA) for small scale preparations and PureLink® HiPure Plasmid Filter Midiprep Kit for larger scale preparations and DNA was quantitated with a Nanodrop® ND-1000 Spectrophotometer (Thermo Fischer Scientific, USA).

    Sequencing:

    Article Title: Phytoplasma SAP11 effector destabilization of TCP transcription factors differentially impact development and defence of Arabidopsis versus maize
    Article Snippet: To generate a plasmid for expression of GFP alone, the ccdB cassette of pUBN-GFP-DEST was replaced with a GFP sequence that carries two translational stop codons instead of the translational start codon. .. The GFP-sequence was amplified from pUBN-GFP-DEST with the gene-specific primers STOP-GFP forward and reverse , cloned into pDONR207 with the Gateway BP Clonase II Enzyme Mix (Invitrogen) and transferred to pUBN-GFP-DEST using the Gateway LR Clonase II Enzyme Mix (Invitrogen).

    Article Title: Cytokine exocytosis and JAK/STAT activation in the Drosophila ovary requires the vesicle trafficking regulator α-Snap
    Article Snippet: The Kozak sequence (CAAC) ( ) was also added to the forward primer just upstream of the start codon. .. The BP reaction was carried out using a pDNOR221 vector (Invitrogen, 12536-017) and Gateway BP Clonase II enzyme mix (Invitrogen, 11789020) followed by heat shock-induced transformation into One Shot OmniMax 2T1 competent E. coli cells (Invitrogen, C854003).

    Article Title: IRAK3 modulates downstream innate immune signalling through its guanylate cyclase activity
    Article Snippet: Generation of IRAK3 and IRAK4 constructs The IRAK3 gene in pBluescriptR (MHS1010-9204142, clone ID 30335802) was cloned into pDONR207 using Gateway® BP Clonase™ II Enzyme Mix (Invitrogen, Thermo Scientific USA). .. Five primers were designed to make four constructs; two forward primers, each containing the Kozak sequence and three reverse primers to either incorporate a Myc tag or a STOP codon (Supplementary Table ) and these were generated by PCR.

    Article Title: Chemical Screening Pipeline for Identification of Specific Plant Autophagy Modulators 1
    Article Snippet: .. The synthetic sequence of attB-flanked SV40 NLS (Eurofins Scientific) was recombined with the Gateway pDONRTM/Zeo vector (ThermoFisher) using Gateway BP Clonase II enzyme mix (ThermoFisher), producing entry clone CC3. .. The CC3 entry clone was then recombined with the AM618 vector to produce an intermediate clone containing the sequence of 2× 35S:: Renilla -NLS fusion::NosT.

    Article Title: Chemical Screening Pipeline for Identification of Specific Plant Autophagy Modulators 1
    Article Snippet: To generate the entry clone AM425, the AtATG8a coding DNA sequence ( ) was amplified from total cDNA of Arabidopsis ( Arabidopsis thaliana ) seedlings using primers AM18/AM19. .. The obtained attB-flanked amplicon was then recombined with the Gateway pDONR/Zeo vector (ThermoFisher) using Gateway BP Clonase II enzyme mix (ThermoFisher).

    Article Title: Phytoplasma SAP11 effector destabilization of TCP transcription factors differentially impact development and defence of Arabidopsis versus maize
    Article Snippet: The cloning of the codon-optimized version of SAP11AYWB without the sequence corresponding to the signal peptide into pDONR207 is described previously [ ]. .. The fragments were further amplified with attB1 and attB2 adapter primers and cloned into pDONR207 with Gateway BP Clonase II Enzyme Mix (Invitrogen, Carlsbad, USA).

    Article Title: Haustoria – arsenals during the interaction between wheat and Puccinia striiformis f. sp. tritici
    Article Snippet: .. For pEDV6, the coding sequence (without a signal peptide) was cloned and recombined in pDONR221 following the instructions of the Gateway BP clonase II enzyme mix and Gateway LR clonase enzyme mix II (Invitrogen, Carlsbad, CA, USA). ..

    Transgenic Assay:

    Article Title: Cytokine exocytosis and JAK/STAT activation in the Drosophila ovary requires the vesicle trafficking regulator α-Snap
    Article Snippet: Paragraph title: Generation of UAS–α-Snap–HA transgenic flies ... The BP reaction was carried out using a pDNOR221 vector (Invitrogen, 12536-017) and Gateway BP Clonase II enzyme mix (Invitrogen, 11789020) followed by heat shock-induced transformation into One Shot OmniMax 2T1 competent E. coli cells (Invitrogen, C854003).

    Article Title: Differential Roles of the Thylakoid Lumenal Deg Protease Homologs in Chloroplast Proteostasis
    Article Snippet: Paragraph title: Generation of Mutant and Transgenic Lines, Genotyping, and Expression Analysis ... The PCR products were recombined with the pDONR221 vector using Gateway BP-Clonase II Enzyme Mix (Invitrogen), yielding entry clones.

    Generated:

    Article Title: Phytoplasma SAP11 effector destabilization of TCP transcription factors differentially impact development and defence of Arabidopsis versus maize
    Article Snippet: The GFP-sequence was amplified from pUBN-GFP-DEST with the gene-specific primers STOP-GFP forward and reverse , cloned into pDONR207 with the Gateway BP Clonase II Enzyme Mix (Invitrogen) and transferred to pUBN-GFP-DEST using the Gateway LR Clonase II Enzyme Mix (Invitrogen). .. Protoplasts were generated from 6-week-old Arabidopsis and four-leaf stage maize plants grown in controlled environmental conditions with a 14h, 22 C°/ 10h, 20°C light / dark period.

    Article Title: IRAK3 modulates downstream innate immune signalling through its guanylate cyclase activity
    Article Snippet: Generation of IRAK3 and IRAK4 constructs The IRAK3 gene in pBluescriptR (MHS1010-9204142, clone ID 30335802) was cloned into pDONR207 using Gateway® BP Clonase™ II Enzyme Mix (Invitrogen, Thermo Scientific USA). .. Five primers were designed to make four constructs; two forward primers, each containing the Kozak sequence and three reverse primers to either incorporate a Myc tag or a STOP codon (Supplementary Table ) and these were generated by PCR.

    Article Title: Differential Roles of the Thylakoid Lumenal Deg Protease Homologs in Chloroplast Proteostasis
    Article Snippet: Transgenic lines expressing His-tagged Deg proteins were generated using directional cloning of PCR products with Gateway technology. .. The PCR products were recombined with the pDONR221 vector using Gateway BP-Clonase II Enzyme Mix (Invitrogen), yielding entry clones.

    Article Title: Phytoplasma SAP11 effector destabilization of TCP transcription factors differentially impact development and defence of Arabidopsis versus maize
    Article Snippet: Generation of Gateway compatible entry clones We generated Gateway compatible entry clones for all experiments, except for the constructs to transform maize. .. The fragments were further amplified with attB1 and attB2 adapter primers and cloned into pDONR207 with Gateway BP Clonase II Enzyme Mix (Invitrogen, Carlsbad, USA).

    Polymerase Chain Reaction:

    Article Title: Host‐induced gene silencing compromises Verticillium wilt in tomato and Arabidopsis
    Article Snippet: Selected DNA fragments were amplified by PCR from the corresponding plasmids using the gene‐specific primers listed in Table S1 (see Supporting Information). .. The DNA fragments were cloned into pDONR207 using the Gateway® BP Clonase® II Enzyme Mix (Invitrogen, Carlsbad, CA, USA) to generate entry vectors, and all the entry vectors were verified by DNA sequencing (Eurofins Genomics, Ebersberg, Germany).

    Article Title: IRAK3 modulates downstream innate immune signalling through its guanylate cyclase activity
    Article Snippet: Generation of IRAK3 and IRAK4 constructs The IRAK3 gene in pBluescriptR (MHS1010-9204142, clone ID 30335802) was cloned into pDONR207 using Gateway® BP Clonase™ II Enzyme Mix (Invitrogen, Thermo Scientific USA). .. Five primers were designed to make four constructs; two forward primers, each containing the Kozak sequence and three reverse primers to either incorporate a Myc tag or a STOP codon (Supplementary Table ) and these were generated by PCR.

    Article Title: Differential Roles of the Thylakoid Lumenal Deg Protease Homologs in Chloroplast Proteostasis
    Article Snippet: .. The PCR products were recombined with the pDONR221 vector using Gateway BP-Clonase II Enzyme Mix (Invitrogen), yielding entry clones. .. Plasmids expressing the Deg genomic DNA with C-terminal His tags were constructed by performing LR-trans reactions between the pDONR221-Deg entry vectors and the pGWB407/pGWB607 destination vectors using the Gateway LR-Clonase II Enzyme Mix (Invitrogen).

    Article Title: A dispensable paralog of succinate dehydrogenase subunit C mediates standing resistance towards a subclass of SDHI fungicides in Zymoseptoria tritici
    Article Snippet: .. To generate the SDHC1 and SDHC3 KO mutants, 5’ upstream regions of 1000bp and 2074 bp and 3’ downstream regions of 914bp and 1313bp for ZtSDHC1 and ZtSDHC3 respectively were PCR-amplified from genomic DNA of IPO323 or 06STD024 strains and the fragments cloned by BP cloning using Gateway BP Clonase II Enzyme Mix (Invitrogen) into pDONR-P4-P1R (upstream regions) or pDONR-P2R-P3 (downstream regions) ( for oligos). .. These 5’ and 3’ gene locus-paired entry plasmids were then combined with the pENTR221-TrpChyg described previously [ ] and pNOV2114_gateway for multisite gateway LR cloning using Gateway LR Clonase II Plus enzyme Mix (Invitrogen) following provider’s instructions.

    Article Title: Phytoplasma SAP11 effector destabilization of TCP transcription factors differentially impact development and defence of Arabidopsis versus maize
    Article Snippet: The full-length ORF of AtTCP6 , AtTCP8 , AtTCP9 , AtTCP12 , AtTCP14 and AtTCP18 ( ) were PCR amplified from complementary DNA (cDNA) with gene-specific primers that contain partial sequences of the attB1 and attB2 Gateway recombination sites ( ). .. The fragments were further amplified with attB1 and attB2 adapter primers and cloned into pDONR207 with Gateway BP Clonase II Enzyme Mix (Invitrogen, Carlsbad, USA).

    Article Title: Complex Iron Uptake by the Putrebactin-Mediated and Feo Systems in Shewanella oneidensis
    Article Snippet: In brief, two fragments flanking the genes of interest were amplified by PCR with outside primers (O; ) containing attB and gene-specific sequences and with inside primers (I; ) containing complementary sequences and gene-specific sequences and were then linked by a second round of PCR with outside primers. .. The fused fragments were introduced into plasmid pHGM01 using Gateway BP clonase II enzyme mix (Invitrogen) according to the manufacturer's instructions.

    Article Title: Haustoria – arsenals during the interaction between wheat and Puccinia striiformis f. sp. tritici
    Article Snippet: The PCR products were ligated into the BSMV:γ vector after digestion with the restriction enzymes Not I and Pac I (Takara, Japan). .. For pEDV6, the coding sequence (without a signal peptide) was cloned and recombined in pDONR221 following the instructions of the Gateway BP clonase II enzyme mix and Gateway LR clonase enzyme mix II (Invitrogen, Carlsbad, CA, USA).

    Infection:

    Article Title: Haustoria – arsenals during the interaction between wheat and Puccinia striiformis f. sp. tritici
    Article Snippet: For pGR106, we amplified HASP candidate genes from the cDNA of Suwon 11 infected with CYR31, digested them with enzymes Cla I, Sal I or Smal I, and cloned into pGR106. .. For pEDV6, the coding sequence (without a signal peptide) was cloned and recombined in pDONR221 following the instructions of the Gateway BP clonase II enzyme mix and Gateway LR clonase enzyme mix II (Invitrogen, Carlsbad, CA, USA).

    Expressing:

    Article Title: Phytoplasma SAP11 effector destabilization of TCP transcription factors differentially impact development and defence of Arabidopsis versus maize
    Article Snippet: Paragraph title: Transient expression assays in Arabidopsis thaliana and maize (Zea mays L.) protoplasts ... The GFP-sequence was amplified from pUBN-GFP-DEST with the gene-specific primers STOP-GFP forward and reverse , cloned into pDONR207 with the Gateway BP Clonase II Enzyme Mix (Invitrogen) and transferred to pUBN-GFP-DEST using the Gateway LR Clonase II Enzyme Mix (Invitrogen).

    Article Title: Host‐induced gene silencing compromises Verticillium wilt in tomato and Arabidopsis
    Article Snippet: The Gateway‐compatible TRV two‐component Agrobacterium ‐mediated expression system was used for gene silencing in tomato, as described previously (Liu et al ., ), and the Gateway‐compatible vectors pFAST R03 (Shimada et al ., ) and pHellsgate 12 (Helliwell and Waterhouse, ) for hairpin RNA‐mediated gene silencing were used to generate stable A. thaliana transformants. .. The DNA fragments were cloned into pDONR207 using the Gateway® BP Clonase® II Enzyme Mix (Invitrogen, Carlsbad, CA, USA) to generate entry vectors, and all the entry vectors were verified by DNA sequencing (Eurofins Genomics, Ebersberg, Germany).

    Article Title: Cytokine exocytosis and JAK/STAT activation in the Drosophila ovary requires the vesicle trafficking regulator α-Snap
    Article Snippet: The coding sequence – excluding the stop codon – to allow expression of the HA tag was amplified using primers that contained attB sites following the GateWay cloning protocol by Invitrogen/ThermoFisher. .. The BP reaction was carried out using a pDNOR221 vector (Invitrogen, 12536-017) and Gateway BP Clonase II enzyme mix (Invitrogen, 11789020) followed by heat shock-induced transformation into One Shot OmniMax 2T1 competent E. coli cells (Invitrogen, C854003).

    Article Title: Differential Roles of the Thylakoid Lumenal Deg Protease Homologs in Chloroplast Proteostasis
    Article Snippet: Paragraph title: Generation of Mutant and Transgenic Lines, Genotyping, and Expression Analysis ... The PCR products were recombined with the pDONR221 vector using Gateway BP-Clonase II Enzyme Mix (Invitrogen), yielding entry clones.

    Article Title: A dispensable paralog of succinate dehydrogenase subunit C mediates standing resistance towards a subclass of SDHI fungicides in Zymoseptoria tritici
    Article Snippet: To generate the SDHC1 and SDHC3 KO mutants, 5’ upstream regions of 1000bp and 2074 bp and 3’ downstream regions of 914bp and 1313bp for ZtSDHC1 and ZtSDHC3 respectively were PCR-amplified from genomic DNA of IPO323 or 06STD024 strains and the fragments cloned by BP cloning using Gateway BP Clonase II Enzyme Mix (Invitrogen) into pDONR-P4-P1R (upstream regions) or pDONR-P2R-P3 (downstream regions) ( for oligos). .. For generating expression constructs under the control of a tetracyclin-repressible promoter, the plasmid pMF2-4h [ ] was modified by removal of the hygromycin resistance cassette after digestion by NotI and recircularization of the plasmid to generate pMF2-4h- .

    Modification:

    Article Title: A dispensable paralog of succinate dehydrogenase subunit C mediates standing resistance towards a subclass of SDHI fungicides in Zymoseptoria tritici
    Article Snippet: To generate the SDHC1 and SDHC3 KO mutants, 5’ upstream regions of 1000bp and 2074 bp and 3’ downstream regions of 914bp and 1313bp for ZtSDHC1 and ZtSDHC3 respectively were PCR-amplified from genomic DNA of IPO323 or 06STD024 strains and the fragments cloned by BP cloning using Gateway BP Clonase II Enzyme Mix (Invitrogen) into pDONR-P4-P1R (upstream regions) or pDONR-P2R-P3 (downstream regions) ( for oligos). .. For generating expression constructs under the control of a tetracyclin-repressible promoter, the plasmid pMF2-4h [ ] was modified by removal of the hygromycin resistance cassette after digestion by NotI and recircularization of the plasmid to generate pMF2-4h- .

    Conjugation Assay:

    Article Title: Complex Iron Uptake by the Putrebactin-Mediated and Feo Systems in Shewanella oneidensis
    Article Snippet: The fused fragments were introduced into plasmid pHGM01 using Gateway BP clonase II enzyme mix (Invitrogen) according to the manufacturer's instructions. .. The resulting vectors were maintained in E. coli DAP auxotroph WM3064 and subsequently transferred into relevant S. oneidensis strains via conjugation.

    Transformation Assay:

    Article Title: Phytoplasma SAP11 effector destabilization of TCP transcription factors differentially impact development and defence of Arabidopsis versus maize
    Article Snippet: The GFP-sequence was amplified from pUBN-GFP-DEST with the gene-specific primers STOP-GFP forward and reverse , cloned into pDONR207 with the Gateway BP Clonase II Enzyme Mix (Invitrogen) and transferred to pUBN-GFP-DEST using the Gateway LR Clonase II Enzyme Mix (Invitrogen). .. Isolation and transformation of Arabidopsis and maize protoplasts were performed as described by [ ].

    Article Title: Host‐induced gene silencing compromises Verticillium wilt in tomato and Arabidopsis
    Article Snippet: The DNA fragments were cloned into pDONR207 using the Gateway® BP Clonase® II Enzyme Mix (Invitrogen, Carlsbad, CA, USA) to generate entry vectors, and all the entry vectors were verified by DNA sequencing (Eurofins Genomics, Ebersberg, Germany). .. All constructs were transformed to Agrobacterium tumefaciens strain GV3101 (pMP90) by electroporation.

    Article Title: Cytokine exocytosis and JAK/STAT activation in the Drosophila ovary requires the vesicle trafficking regulator α-Snap
    Article Snippet: .. The BP reaction was carried out using a pDNOR221 vector (Invitrogen, 12536-017) and Gateway BP Clonase II enzyme mix (Invitrogen, 11789020) followed by heat shock-induced transformation into One Shot OmniMax 2T1 competent E. coli cells (Invitrogen, C854003). .. Successful cloning of α-Snap into pDONR221 was confirmed by sequencing using M13F(-21) and M13R primers (Genewiz).

    Article Title: IRAK3 modulates downstream innate immune signalling through its guanylate cyclase activity
    Article Snippet: Generation of IRAK3 and IRAK4 constructs The IRAK3 gene in pBluescriptR (MHS1010-9204142, clone ID 30335802) was cloned into pDONR207 using Gateway® BP Clonase™ II Enzyme Mix (Invitrogen, Thermo Scientific USA). .. Heat shock transformation was used for all One Shot® Max Efficiency® DH5α™-T1R competent cells and bacteria were grown on LB plates or liquid broth containing the appropriate antibiotic (e.g. ampicillin 100 μg/ml or gentamycin 15 μg/ml).

    Article Title: A dispensable paralog of succinate dehydrogenase subunit C mediates standing resistance towards a subclass of SDHI fungicides in Zymoseptoria tritici
    Article Snippet: Production of Z . tritici transformants The multisite binary pNOV2114_gateway and pNOV2114 Hyg _gateway (3-way) vectors were used to generate the different transformation constructs [ ]. .. To generate the SDHC1 and SDHC3 KO mutants, 5’ upstream regions of 1000bp and 2074 bp and 3’ downstream regions of 914bp and 1313bp for ZtSDHC1 and ZtSDHC3 respectively were PCR-amplified from genomic DNA of IPO323 or 06STD024 strains and the fragments cloned by BP cloning using Gateway BP Clonase II Enzyme Mix (Invitrogen) into pDONR-P4-P1R (upstream regions) or pDONR-P2R-P3 (downstream regions) ( for oligos).

    Spectrophotometry:

    Article Title: IRAK3 modulates downstream innate immune signalling through its guanylate cyclase activity
    Article Snippet: Generation of IRAK3 and IRAK4 constructs The IRAK3 gene in pBluescriptR (MHS1010-9204142, clone ID 30335802) was cloned into pDONR207 using Gateway® BP Clonase™ II Enzyme Mix (Invitrogen, Thermo Scientific USA). .. Plasmids were purified using PureLink® Quick Plasmid Miniprep Kit (Thermo Fisher Scientific, USA) for small scale preparations and PureLink® HiPure Plasmid Filter Midiprep Kit for larger scale preparations and DNA was quantitated with a Nanodrop® ND-1000 Spectrophotometer (Thermo Fischer Scientific, USA).

    Plasmid Preparation:

    Article Title: Phytoplasma SAP11 effector destabilization of TCP transcription factors differentially impact development and defence of Arabidopsis versus maize
    Article Snippet: To generate a plasmid for expression of GFP alone, the ccdB cassette of pUBN-GFP-DEST was replaced with a GFP sequence that carries two translational stop codons instead of the translational start codon. .. The GFP-sequence was amplified from pUBN-GFP-DEST with the gene-specific primers STOP-GFP forward and reverse , cloned into pDONR207 with the Gateway BP Clonase II Enzyme Mix (Invitrogen) and transferred to pUBN-GFP-DEST using the Gateway LR Clonase II Enzyme Mix (Invitrogen).

    Article Title: Host‐induced gene silencing compromises Verticillium wilt in tomato and Arabidopsis
    Article Snippet: The DNA fragments were cloned into pDONR207 using the Gateway® BP Clonase® II Enzyme Mix (Invitrogen, Carlsbad, CA, USA) to generate entry vectors, and all the entry vectors were verified by DNA sequencing (Eurofins Genomics, Ebersberg, Germany). .. Subsequently, the entry vector pDONR207 carrying the Ave1 fragment was transferred to TRV2 and pFAST R03 to generate constructs TRV2::Ave1 and pFAST R03_Ave1 (Fig. ), respectively, and pDONR207 entry vectors carrying the NLP1 or Sge1 fragment were recombined into pTRV2 and pHellsgate 12 to generate constructs TRV2::NLP1 , TRV2::Sge1 , pHellsgate 12_NLP1 and pHellsgate 12_Sge1 (Fig. ) using Gateway® LR Clonase® II Enzyme Mix (Invitrogen).

    Article Title: Cytokine exocytosis and JAK/STAT activation in the Drosophila ovary requires the vesicle trafficking regulator α-Snap
    Article Snippet: .. The BP reaction was carried out using a pDNOR221 vector (Invitrogen, 12536-017) and Gateway BP Clonase II enzyme mix (Invitrogen, 11789020) followed by heat shock-induced transformation into One Shot OmniMax 2T1 competent E. coli cells (Invitrogen, C854003). .. Successful cloning of α-Snap into pDONR221 was confirmed by sequencing using M13F(-21) and M13R primers (Genewiz).

    Article Title: IRAK3 modulates downstream innate immune signalling through its guanylate cyclase activity
    Article Snippet: Generation of IRAK3 and IRAK4 constructs The IRAK3 gene in pBluescriptR (MHS1010-9204142, clone ID 30335802) was cloned into pDONR207 using Gateway® BP Clonase™ II Enzyme Mix (Invitrogen, Thermo Scientific USA). .. Plasmids were purified using PureLink® Quick Plasmid Miniprep Kit (Thermo Fisher Scientific, USA) for small scale preparations and PureLink® HiPure Plasmid Filter Midiprep Kit for larger scale preparations and DNA was quantitated with a Nanodrop® ND-1000 Spectrophotometer (Thermo Fischer Scientific, USA).

    Article Title: Chemical Screening Pipeline for Identification of Specific Plant Autophagy Modulators 1
    Article Snippet: .. The synthetic sequence of attB-flanked SV40 NLS (Eurofins Scientific) was recombined with the Gateway pDONRTM/Zeo vector (ThermoFisher) using Gateway BP Clonase II enzyme mix (ThermoFisher), producing entry clone CC3. .. The CC3 entry clone was then recombined with the AM618 vector to produce an intermediate clone containing the sequence of 2× 35S:: Renilla -NLS fusion::NosT.

    Article Title: Differential Roles of the Thylakoid Lumenal Deg Protease Homologs in Chloroplast Proteostasis
    Article Snippet: .. The PCR products were recombined with the pDONR221 vector using Gateway BP-Clonase II Enzyme Mix (Invitrogen), yielding entry clones. .. Plasmids expressing the Deg genomic DNA with C-terminal His tags were constructed by performing LR-trans reactions between the pDONR221-Deg entry vectors and the pGWB407/pGWB607 destination vectors using the Gateway LR-Clonase II Enzyme Mix (Invitrogen).

    Article Title: Chemical Screening Pipeline for Identification of Specific Plant Autophagy Modulators 1
    Article Snippet: .. The obtained attB-flanked amplicon was then recombined with the Gateway pDONR/Zeo vector (ThermoFisher) using Gateway BP Clonase II enzyme mix (ThermoFisher). .. Fluc and Rluc coding sequences were amplified using the primer pairs PB31/PB32 and PB33/PB34 and vectors pRD400 35S::Fluc and pRD400 35S::Rluc , respectively.

    Article Title: A dispensable paralog of succinate dehydrogenase subunit C mediates standing resistance towards a subclass of SDHI fungicides in Zymoseptoria tritici
    Article Snippet: To generate the SDHC1 and SDHC3 KO mutants, 5’ upstream regions of 1000bp and 2074 bp and 3’ downstream regions of 914bp and 1313bp for ZtSDHC1 and ZtSDHC3 respectively were PCR-amplified from genomic DNA of IPO323 or 06STD024 strains and the fragments cloned by BP cloning using Gateway BP Clonase II Enzyme Mix (Invitrogen) into pDONR-P4-P1R (upstream regions) or pDONR-P2R-P3 (downstream regions) ( for oligos). .. For generating expression constructs under the control of a tetracyclin-repressible promoter, the plasmid pMF2-4h [ ] was modified by removal of the hygromycin resistance cassette after digestion by NotI and recircularization of the plasmid to generate pMF2-4h- .

    Article Title: Complex Iron Uptake by the Putrebactin-Mediated and Feo Systems in Shewanella oneidensis
    Article Snippet: .. The fused fragments were introduced into plasmid pHGM01 using Gateway BP clonase II enzyme mix (Invitrogen) according to the manufacturer's instructions. .. The resulting vectors were maintained in E. coli DAP auxotroph WM3064 and subsequently transferred into relevant S. oneidensis strains via conjugation.

    Article Title: Haustoria – arsenals during the interaction between wheat and Puccinia striiformis f. sp. tritici
    Article Snippet: The PCR products were ligated into the BSMV:γ vector after digestion with the restriction enzymes Not I and Pac I (Takara, Japan). .. For pEDV6, the coding sequence (without a signal peptide) was cloned and recombined in pDONR221 following the instructions of the Gateway BP clonase II enzyme mix and Gateway LR clonase enzyme mix II (Invitrogen, Carlsbad, CA, USA).

    DNA Sequencing:

    Article Title: Host‐induced gene silencing compromises Verticillium wilt in tomato and Arabidopsis
    Article Snippet: .. The DNA fragments were cloned into pDONR207 using the Gateway® BP Clonase® II Enzyme Mix (Invitrogen, Carlsbad, CA, USA) to generate entry vectors, and all the entry vectors were verified by DNA sequencing (Eurofins Genomics, Ebersberg, Germany). .. Subsequently, the entry vector pDONR207 carrying the Ave1 fragment was transferred to TRV2 and pFAST R03 to generate constructs TRV2::Ave1 and pFAST R03_Ave1 (Fig. ), respectively, and pDONR207 entry vectors carrying the NLP1 or Sge1 fragment were recombined into pTRV2 and pHellsgate 12 to generate constructs TRV2::NLP1 , TRV2::Sge1 , pHellsgate 12_NLP1 and pHellsgate 12_Sge1 (Fig. ) using Gateway® LR Clonase® II Enzyme Mix (Invitrogen).

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    Thermo Fisher gateway bp clonase ii enzyme mix
    Gateway Bp Clonase Ii Enzyme Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 155 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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